What is malachite green staining?
The Malachite green staining ( Schaeffer-Fulton method ) is the most common method used to perform endospore staining. Malachite green stain can also used as a simple stain for bacterial cells. The Schaeffer-Fulton method uses heat to push the primary dye (malachite green) into the endospore.
What diseases can malachite green be used to treat?
Disorders treated by Malachite Green include the following: Egg Fungus - Serves as a safe disinfectant for fish eggs. Ichthyophthirius multi files - Small white spots resembling sand, fish scratches against objects. Oödinium pillars - Also known as Velvet, this is actually caused by a parasite.
What is Leuco malachite green?
The leuco form of malachite green was first prepared by Hermann Fischer in 1877 by condensing benzaldehyde and dimethylaniline in the molecular ratio 1:2 in the presence of sulfuric acid. Malachite green is traditionally used as a dye.
What is the LD50 of malachite green?
The LD 50 (oral, mouse) is 80 mg/kg. Rats fed malachite green experience "a dose-related increase in liver DNA adducts" along with lung adenomas. Leucomalachite green causes an "increase in the number and severity of changes".
What is malachite green used for?
Malachite green is an organic chloride salt that is the monochloride salt of malachite green cation. Used as a green-coloured dye, as a counter-stain in histology, and for its anti-fungal properties in aquaculture.
How does malachite green stain work?
Malachite green is water soluble and has a low affinity for cellular material, so vegetative cells may be decolorized with water. Safranin is then applied to counterstain any cells which have been decolorized. At the end of the staining process, vegetative cells will be pink, and endospores will be dark green.
What type of dye is malachite green?
cationic dyeMalachite green (MG) is water soluble cationic dye that appears as green crystalline powder and belongs to triphenylmethane category (Raval et al. 2016b).
Is malachite green a mordant?
Thus it is found that malachite green is direct dye for wool but is a mordant dye for cotton.
Which stain is used for spore?
Malachite GreenThe bacterial endospores need a staining which can penetrate wall thickness of spore bacteria. A method of endospores staining is Schaeffer Fulton method that used Malachite Green. It is an alkaline substance staining that can staining the spore bacteria.
Why is it important to leave the malachite green on the slide for at least 10 minutes in the endospore staining procedure?
Why is it important to leave the malachite green on the slide for at least 10 minutes in the endospore staining procedure? In using the Schaeffer-Fulton Method, the without heat method, to view endospores you must leave stain on for a long time to allow it to penetrate the spore coat.
Is malachite green a basic stain?
Malachite Green 1% w/v is used as staining solution in spore staining and simple staining. Malachite Green is used for bacterial spore staining by Schaeffer and Fulton's method. It can also be used as a simple stain for bacterial cells and in place of methyl-green in Pappenheim stain, when combined with Gram stain.
Is malachite green toxic to humans?
The toxicity of this dye increases with exposure time, temperature and concentration. It has been reported to cause carcinogenesis, mutagenesis, chromosomal fractures, teratogenecity and respiratory toxicity.
How do you make malachite green dye?
Malachite green can be prepared by reaction of Benzaldehyde with N, N-dimethyl aniline in the presence of H₂SO₄ to form a triphenylmethane derivative. The subsequent oxidation with PbO₂ followed by treatment with an excess of concentrated HCl yields a green dye known as malachite green.
Is malachite green Banned?
Although malachite green (MG), is banned in Europe and US for its carcinogenic and teratogenic effect, the dye being cheap, is persistently used in various countries for fish farming, silk, dye, leather and textile industries.
Is malachite green sensitive to light?
Malachite Green Reagent is light sensitive so using dark opaque tubes or wrapping tubes in foil is recommended.
What is mordant green dye?
A mordant or dye fixative is a substance used to set (i.e. bind) dyes on fabrics by forming a coordination complex with the dye, which then attaches to the fabric (or tissue). It may be used for dyeing fabrics or for intensifying stains in cell or tissue preparations.
What is malachite green for Endospore staining?
Malachite green is forced into the spores by steaming the bacterial emulsion. This heating step stains the vegetative cells and the endospores. The water acts as a decolorizer for the vegetative cells, but the stain is not released by the endospores and free spores.
How do you make a malachite green indicator?
Preparation. The leuco form of malachite green was first prepared by Hermann Fischer in 1877 by condensing benzaldehyde and dimethylaniline in the molecular ratio 1:2 in the presence of sulfuric acid.
Why does malachite have a blue green colour?
This geological mineral is made of a compound called copper carbonate, which takes on various shades of blue to green. However, the therapeutic agent malachite green contains zero copper, and the only similarity between the mineral and medicinal ingredient is in name and colour.
Is malachite green sensitive to light?
Malachite Green Reagent is light sensitive so using dark opaque tubes or wrapping tubes in foil is recommended.
Method 1 (Short Term Bath)
1. Change the carbon in the outside filter or add an outside filter with fresh carbon to the hatchery tank. Use a high-quality liquid phase activat...
Method 2 (Dip)
1. To a clean, non-metallic container, add 1 gallon of aged hatchery water, or water treated with NovAqua (DFS# 21207, 21208). 2. Add 5 Fl. oz. of...
Treatment For Fungus on Fishes
1. Transfer fishes to a separate quarantine tank, if possible. 2. Remove carbon, clean outside filter and return to use with clean mechanical filte...
What is malachite green used for?
Malachite green is traditionally used as a dye for materials such as silk, leather, and paper. Despite its name the dye is not prepared from the mineral malachite, and the name just comes from the similarity of color.
When was malachite green first made?
The leuco form of malachite green was first prepared by Hermann Fischer in 1877 by condensing benzaldehyde and dimethylaniline in the molecular ratio 1:2 in the presence of sulfuric acid.
What does the green color mean in blood?
Leuco-malachite green (LMG) is used as a detection method for latent blood in forensic science. Hemoglobin catalyzes the reaction between LMG and hydrogen peroxide, converting the colorless LMG into malachite green. Therefore, the appearance of a green color indicates the presence of blood.
What is LMG in fish?
The principal metabolite, LMG, is found in fish treated with malachite green, and this finding is the basis of controversy and government regulation. See also Antimicrobials in aquaculture . MG has frequently been used to catch thieves and pilferers. The bait, usually money, is sprinkled with the anhydrous powder.
What is the organic dye called?
This article is about the organic dye called "malachite green". For the use of malachite as a pigment, see Malachite § Use, and Basic copper carbonate § Uses.
Is malachite green safe to eat?
In 1992, Canadian authorities determined that eating fish contaminated with malachite green posed a significant health risk. Malachite green was classified a Class II Health Hazard. Due to its low manufacturing cost, malachite green is still used in certain countries with less restrictive laws for non aquaculture purposes. In 2005, analysts in Hong Kong found traces of malachite green in eels and fish imported from China. In 2006, the United States Food and Drug Administration (FDA) detected malachite green in seafood imported from China, among others, where the substance is also banned for use in aquaculture. In June 2007, the FDA blocked the importation of several varieties of seafood due to continued malachite green contamination. The substance has been banned in the United States since 1983 in food-related applications. It is banned in the UK, also. Animals metabolize malachite green to its leuco form. Being lipophillic (the leuco form has a log P of 5.70), the metabolite is retained in catfish muscle longer ( HL = 10 days) than is the parent molecule (HL = 2.8 days).
Does malachite green have fluorescence?
Although malachite green has almost no fluorescence in aqueous solution ( quantum yield 7.9x10 −5 ), several research groups have developed technologies to detect malachite green. For example, Zhao et al., demonstrated the use of malachite green aptamer in microcantilever based sensors to detect low concentration of malachite green.
What is malachite green used for?
History & Uses: First made in 1877, Malachite Green has been used primarily as a dye for fabrics, leather, and paper. It is also used as a biological stain. In aquaculture, it is often used as an anti-fungal for eggs in hatcheries, in spite of growing concern regarding potential health hazards.
How long does it take for Malachite Green to go away from water?
Allow the filter to run and allow it to remove the Malachite Green from the water for one hour. Note: If any color remains in the water after 1 hour, perform a partial water change of at least 25%, change filter carbon and allow it to remove any additional Malachite Green.
How much Kordon Malachite Green is in 5 oz?
Add 5 Fl. oz. of 0.038% Kordon Malachite Green (approximately 149 ml). This produces a concentration of 1500 ppm.
What is Shirlie's job?
Shirlie is a fish and aquarium lover with 16 years of experience writing on the topic of raising and keeping fish at home. Action: Triarulmaethane dye, antibacterial, antifungal, anti-parasitic. History & Uses: First made in 1877, Malachite Green has been used primarily as a dye for fabrics, leather, and paper.
How long does it take for ich to cure?
Remove and discard all carbon from filter, but DO NOT discontinue filtration. Typically cures ich in 24 hours, treatment can be repeated in 24-48 hours, if necessary. If fish experience stress, change 50% of the water.
Is Malachite Green the same as Methylene Blue?
Warning. Malachite Green is often combined with formaldehyde (aka formalin) and is somewhat similar to Methylene Blue in terms of uses. However, it is a harsher chemical and must be used with caution. It is most effective against external parasites, particularly when combined with formaldehyde.
What is the most widely used stain technique for endospores?
Schaeffer-Fulton stain technique. It is the most widely used technique for endospore staining. The technique was first described by Alice B. Schaeffer and MacDonald Fulton in the 1930s. The method utilizes malachite green as the primary stain and safranin as counterstain.
How to stain endospores?
Prepare smears of organisms to be tested for the presence of endospores on a clean microscope slide and air dry it. Heat fix the smear. Place a small piece of blotting paper (absorbent paper) over the smear and place the slide (smear side up) on a wire gauze on a ring stand.
What happens when you decolorize with acid alcohol?
When decolorized with acid alcohol color washes off the vegetative cells and makes them colorless. Since the counterstain nigrosin is negatively charged, bacterial cells don’t easily take up the counterstain. Therefore, vegetative cells appear colorless, endospores stain red, and the background is black.
What is endospore staining?
Last updated on June 21st, 2021. Endospore staining is a differential staining technique that selectively stains the spores and makes them distinguishable from the vegetative part of the cells. Endospores are produced by a few genera of Gram-positive bacilli such as Bacillus and Clostridium, in response to adverse environmental conditions.
How to remove secondary stain from slide?
Rinse both sides of the slide to remove the secondary stain and blot the slide/ air dry.
Is endospore stain the same as counterstain?
Although the principle of endospore staining is the same, variations exist in the choice of primary stain, counterstain, and whether or not decolorizer is used. Some of them are summarized below
Can endospores be seen on Gram stain?
Spores can generally be recognized on Gram’s stains (endospores do not stain and appear as refractile, nonstaining bodies). Endospores can also be demonstrated in unstained wet films under a phase-contrast microscope. They appear as large refractile oval or spherical bodies within the mother cell.
General Properties of Malachite Green
- The structure of malachite green is given as follows: Imagewillbeuploadedsoon Further, the green coloured compound is given as : Imagewillbeuploadedsoon Malachite green is a chemical compound that is generally classified in the dyestuff industry. It is used as a triarylmethane dye and also extensively used in the pigment industry. In scientific ter...
Preparation of Malachite Green
- Hermann Fischer was the first one to prepare Leuco Malachite Green in 1877. He did so by condensing the benzaldehyde and dimethylaniline in the molecular ratio of 1:2. The reaction was carried out in the presence of sulphuric acid. The complete reaction is given below: Imagewillbeuploadedsoon
Detection and Uses of Malachite Green
- Malachite green is hard to detect. This is because it does not give out any fluorescence in an aqueous solution, a common method to detect fluorescence. Many research groups have developed different methods and techniques to detect malachite green. An example of this is the use of malachite green aptamer in microcantilever based sensors by Zhao et al. It is used to det…
Toxicity & Regulation of Malachite Green
- In biological experimentation, rats are fed the malachite green drug LD50. The ones that have been fed with malachite green, experience an increase in liver DNA along with the lung adenomas. Leuco Malachite Green is also observed to cause an increase in the severity of the changes. As the LMG is the primary metabolite of malachite green found retained in the fish mu…
Overview
Malachite green is an organic compound that is used as a dyestuff and controversially as an antimicrobial in aquaculture. Malachite green is traditionally used as a dye for materials such as silk, leather, and paper. Despite its name the dye is not prepared from the mineral malachite; the name just comes from the similarity of color.
Uses
Malachite green is traditionally used as a dye. Kilotonnes of MG and related triarylmethane dyes are produced annually for this purpose.
MG is active against the oomycete Saprolegnia, which infects fish eggs in commercial aquaculture, MG has been used to treat Saprolegnia and is used as an antibacterial. It is a very popular treatment against Ichthyophthirius multifiliis in
Structures and properties
Malachite green is classified in the dyestuff industry as a triarylmethane dye and also using in pigment industry. Formally, malachite green refers to the chloride salt [C6H5C(C6H4N(CH3)2)2]Cl, although the term malachite green is used loosely and often just refers to the colored cation. The oxalate salt is also marketed. The anions have no effect on the color. The intense green color of th…
Preparation
The leuco form of malachite green was first prepared by Hermann Fischer in 1877 by condensing benzaldehyde and dimethylaniline in the molecular ratio 1:2 in the presence of sulfuric acid.
Regulation
In 1992, Canadian authorities determined that eating fish contaminated with malachite green posed a significant health risk. Malachite green was classified a Class II Health Hazard. Due to its low manufacturing cost, malachite green is still used in certain countries with less restrictive laws for non aquaculture purposes. In 2005, analysts in Hong Kong found traces of malachite green in eels and fish imported from China. In 2006, the United States Food and Drug Administration (FD…
Toxicity
The LD50 (oral, mouse) is 80 mg/kg. Rats fed malachite green experience "a dose-related increase in liver DNA adducts" along with lung adenomas. Leucomalachite green causes an "increase in the number and severity of changes". As leucomalachite green is the primary metabolite of malachite green and is retained in fish muscle much longer, most human dietary intake of malachite green from eating fish would be in the leuco form. During the experiment, rats were fed up to 543 ppm …
Detection
Although malachite green has almost no fluorescence in aqueous solution (quantum yield 7.9x10 ), several research groups have developed technologies to detect malachite green. For example, Zhao et al., demonstrated the use of malachite green aptamer in microcantilever based sensors to detect low concentration of malachite green.
Further reading
• Cho, Bongsup P.; Yang, Tianle; Blankenship, Lonnie R.; et al. (2003). "Synthesis and Characterization of N-Demethylated Metabolites of Malachite Green and Leucomalachite Green". Chem. Res. Toxicol. 16 (3): 285–294. doi:10.1021/tx0256679. PMID 12641428.
• Plakas, S. M.; El Said, K. R.; Stehly, G. R.; et al. (1996). "Uptake, tissue distribution, and metabolism of malachite green in the channel catfish (Ictalurus punctatus)". Can. J. Fish. Aquat. Sci. 53 (6): 1427–1433. doi:10.1139/cjfas …